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Cloning and characterization of 2-C-methyl-d-erythritol-4-phosphate pathway genes for isoprenoid biosynthesis from Indian ginseng, Withania somnifera

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Abstract

Withania somnifera (L.) is one of the most valuable medicinal plants used in Ayurvedic and other indigenous medicines. Pharmaceutical activities of this herb are associated with presence of secondary metabolites known as withanolides, a class of phytosteroids synthesized via mevalonate (MVA) and 2-C-methyl-d-erythritol-4-phosphate pathways. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized two genes encoding 1-deoxy-d-xylulose-5-phosphate synthase (DXS; EC 2.2.1.7) and 1-deoxy-d-xylulose-5-phosphate reductase (DXR; EC 1.1.1.267) enzymes involved in the biosynthesis of isoprenoids. The full-length cDNAs of W. somnifera DXS (WsDXS) and DXR (WsDXR) of 2,154 and 1,428 bps encode polypeptides of 717 and 475 amino acids residues, respectively. The expression analysis suggests that WsDXS and WsDXR are differentially expressed in different tissues (with maximal expression in flower and young leaf), chemotypes of Withania, and in response to salicylic acid, methyl jasmonate, as well as in mechanical injury. Analysis of genomic organization of WsDXS shows close similarity with tomato DXS in terms of exon–intron arrangements. This is the first report on characterization of isoprenoid biosynthesis pathway genes from Withania.

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Abbreviations

DMAPP:

Dimethylallyl diphosphate

DXP:

1-Deoxy-d-xylulose-5-phosphate

DXR:

1-Deoxy-d-xylulose-5-phosphate reductase

DXS:

1-Deoxy-d-xylulose-5-phosphate synthase

IPP:

Isoprenoid diphosphate

MeJA:

Methyl jasmonate

MEP pathway:

2-C-Methyl-d-erythritol-4-phosphate pathway

MVA pathway:

Mevalonate pathway

SA:

Salicylic acid

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Acknowledgments

The authors are thankful to the Council of Scientific and Industrial Research, New Delhi, Govt. of India, for providing the financial support to carry out this work under NMITLI scheme and Network Project (NWP-08). Authors acknowledge Dr. Satya Narain Jena, Scientist, CSIR-NBRI, for helping in sequencing of different clones used in the study.

Conflict of interest

All the authors declare that we have no conflict of interest.

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Authors and Affiliations

  1. National Botanical Research Institute, Council of Scientific and Industrial Research, Rana Pratap Marg, Lucknow, 226001, India

    Parul Gupta, Aditya Vikram Agarwal, Nehal Akhtar & Prabodh Kumar Trivedi

  2. Central Institute of Medicinal and Aromatic Plants, Council of Scientific and Industrial Research, Lucknow, 226015, India

    Rajender Singh Sangwan

  3. Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi, 221005, India

    Surya Pratap Singh

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  1. Parul Gupta
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  2. Aditya Vikram Agarwal
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  3. Nehal Akhtar
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  4. Rajender Singh Sangwan
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  5. Surya Pratap Singh
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  6. Prabodh Kumar Trivedi
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Correspondence to Prabodh Kumar Trivedi.

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Gupta, P., Agarwal, A.V., Akhtar, N. et al. Cloning and characterization of 2-C-methyl-d-erythritol-4-phosphate pathway genes for isoprenoid biosynthesis from Indian ginseng, Withania somnifera . Protoplasma 250, 285–295 (2013). https://doi.org/10.1007/s00709-012-0410-x

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